Molecular and classical chromosomal techniques reveal diversity in bushcricket genera of Barbitistini (Orthoptera).

The cytogenetic characteristics of 17 species of bushcricket belonging to eight genera of the tribe Barbitistini were examined by fluorescence in situ hybridization with 18S rDNA and (TTAGGn) telomeric as probes and by C-banding, silver, and fluorochrome staining. These markers were used to understand chromosomal organization and evolutionary relationships between genera or species within the same genus. The number of 18S rDNA clusters per haploid genome that co-localized with active nucleolus organizer regions (NORs) ranged from one to five, with the most common pattern being the presence of one NOR-bearing chromosome. This ribosomal cistron was preferentially located in the paracentromeric region of autosomes and very rarely in the sex chromosome. The results demonstrated coincidence between the localization of major ribosomal genes and active NORs and the position of C-band and GC-rich regions. The rDNA/NOR distribution and the composition of chromosome heterochromatin proved to be good cytogenetic markers for distinguishing species and phylogenetic lines and for understanding the genomic differentiation and evolution of Barbitistini. A comparison of cytogenetic and morphological or behavioral traits suggests that morphological and behavioral specialization in this group was not followed by major karyotype modification (except for Leptophyes). However, the occurrence and distribution of different repetitive DNA sites tends to vary among the taxa.

2-Oxoglutaric acid administration diminishes fundectomy-induced osteopenia in pigs.

Partial surgical removal of the stomach (fundectomy, FX) leads to osteopenia in animals and humans. FX adversely affects the bone. 2-oxoglutaric acid is a precursor of glutamine and hydroxyproline--the most abundant amino acid of collagen. The aim of the study was to investigate the effects of 2-oxoglutaric acid on FX-evoked osteopenia in pigs. Eighteen castrated male pigs of the Pulawska breed were used. Twelve pigs were subjected to FX and divided into two groups: FX + AKG (the AKG group; AKG at the daily dosage of 0.4 g/kg of body weight) and FX + Placebo (the FXC group; received CaCO(3) as placebo). Remaining six pigs were sham-operated (the SHO group). The pigs were euthanized at the age of 8 months and long bones were collected. Area bone mineral density (aBMD) and bone mineral content (BMC) were measured; morphology, geometry and biomechanical properties were determined. Moreover, the serum concentrations of selected hormones and one marker of bone metabolism were determined. FX caused osteopenia in the pigs and treatment with AKG greatly reduced these effects of FX in pigs. Negative effect of fundectomy on the skeletal system leading to decreased bone mass in pigs is associated with lowered body gain and activity of the gastric-hypothalamic-pituitary axis. Better definitions of each of the local and systemic hormonal and structural components associated with fundectomy-induced decreased bone mass that separately and together determine the whole bone properties may lead to identify opportunities for prevention.

Bone development of suckling piglets after prenatal, neonatal or perinatal treatment with dexamethasone.

In mammals, the release from growth-inhibiting conditions results in catch-up growth. To investigate animal evidence for whether prenatal dexamethasone (DEX) treatment leads to the development of growth restriction especially reduced mineralization of skeleton, and release from it leads to the phenomenon of catch-up, piglets were prenatally exposed to DEX (3.0 mg/sow per day(-2)) during the last 24 days of prenatal life and tested further in two different ways: discontinued at birth and continued administration of DEX (0.5 mg/kg day(-2)) to piglets through 30 days of neonatal life. Using dual energy X-ray absorptiometry methods, bone mineral density (BMD) and bone mineral content (BMC) were measured. The three-point bending test was applied to determine the mechanical properties of the bones. Furthermore, geometric properties of the bones were assessed. Serum concentration of osteocalcin (OC) was determined. Histomorphological analysis of the ribs was also performed. The consequences of neonate DEX treatment and in utero DEX exposure were reflected in a dramatic decrease of BMD, BMC and blood serum OC concentration and geometric parameters of piglets' bones. Prenatal action of DEX during the last 24 days of pregnancy resulted in continued neonatal modification of bone tissues, thus diminishing bone quality, and negatively influenced structural development and mechanical properties, finally increasing the risk of fractures of ribs and limb bones. Prenatal DEX treatment limited to the last 24 days of foetal life did not reduce the term birth weight and the growth of suckling piglets followed up to 30 days of neonatal life, and catch-up in bone mineralization did not occur.

Alpha-ketoglutarate protects the liver of piglets exposed during prenatal life to chronic excess of dexamethasone from metabolic and structural changes.

Glucocorticoids play a role in the origin of the features of the metabolic diseases. Alpha-ketoglutarate (AKG) is defined as glutamine homologue and derivative, conditionally an essential amino acid. In the liver, glutamine serves as a precursor for ureagenesis, gluconeogenesis and acute phase protein synthesis The aim of the study was to determine the effect of AKG administered to piglets prenatally exposed to dexamethasone, on the structure of the liver and its metabolic function. Sows were administered with dexamethasone (3 mg/sow/48 h) from day 70 of pregnancy to the parturition, and then after the birth, the piglets were divided into the group administered with AKG (0.4 g/kg body weight) or physiological saline. Biochemical markers, lysozyme and ceruloplasmin serum activities, concentrations of selected free amino acids, macro- and microelements and histomorphometry of the liver tissue were determined. The total cholesterol concentrations in the sows and their newborns from the Dex groups were higher by 72% and 64%, respectively, compared with the control groups. Triacylglycerol concentration was higher by 50% in sows from the Dex group and 55% in the new-born piglets. Alpha-ketoglutarate administered to the piglets after prenatal influence of dexamethasone lowered the total cholesterol concentration by 40%, and enhanced aspartate by 41%, serine by 76%, glutamate by 105%, glutamine by 36%, glycine by 53% and arginine by 105%, as well as methionine and cystathionine, but increased the sulphur concentration compared with the control (p < 0.01). Intracellular space D decreased after AKG administration in comparison with the piglets from Dex/Control group not treated with AKG. Postnatal administration of AKG had a protective effect on liver structure, and lowered the total cholesterol concentration in piglets prenatally exposed to dexamethasone, and also influenced selected macro- and microelement serum concentrations and amino acids plasma concentration.

Effect of maternal dexamethasone and alpha-ketoglutarate administration on skeletal development during the last three weeks of prenatal life in pigs.

BACKGROUND: The effect of dexamethasone (Dex) on postnatal bone formation processes is known to decrease the synthesis of collagen and bone matrix, but the effect of alpha-ketoglutarate (AKG) is to induce positive effects on growth and skeletal development during postnatal life. However, the effects of Dex and AKG treatment on the prenatal processes of skeletal development have not been investigated so far. OBJECTIVE: The aim of this study was to determine the effect of Dex and AKG administered separately or simultaneously to sows during the last three weeks of pregnancy on the skeletal development in fetuses. METHODS: Immediately after birth blood samples were collected from non-suckling piglets for alkaline phosphatase and osteocalcin determinations, and the humeri were isolated. Bone mineral density (BMD) and bone mineral content (BMC) of humeri and the geometric and mechanical properties were evaluated. RESULTS: Dex and AKG administered separately to pregnant sows during the last 24 days of prenatal life decreased BMD, BMC, and geometric and mechanical parameters of humeri in the newborns. Simultaneous administration of Dex and AKG significantly increased the analyzed properties of humeri. CONCLUSION: The bone mineral density and mechanical and geometric properties of humeri indicate an inverse effect of maternal separate or simultaneous administration of AKG and Dex to sows on bone development during the last 24 days of prenatal life.

Effects of alpha-ketoglutarate on bone homeostasis and plasma amino acids in turkeys.

The objective of the study was to evaluate the effect of denervation and alpha-ketoglutarate (AKG) administration on the development of osteopenia in the turkey radius. At 22 d of age, all turkeys were subjected to neurectomy of the right radius. Control turkeys were given a saline solution into the crop each day for 97 d. Experimental turkeys were given 0.4 g of AKG/kg of BW into the crop each day. After 98 d, BW was not affected by the AKG treatment. Volumetric bone mineral density of the radius was measured by quantitative computed tomography. Mechanical properties were tested using a 3-point bending test. Cross-sectional area, second moment of inertia, and mean relative wall thickness were measured as well. Amino acid concentrations were assessed with the use of ion-exchange chromatography. Denervation had a negative effect on all bone characteristics that were measured except bone length. The AKG had a positive effect on all bone characteristics except bone length. Plasma concentrations of proline and leucine were increased by AKG, whereas concentrations of taurine and glutamine were decreased. The turkey radius appears to be a good model for studying osteopenia because its development can be affected by treatments such as denervation and AKG administration.

Heavy metal accumulation, heat shock protein expression and cytogenetic changes in Tetrix tenuicornis (L.) (Tetrigidae, Orthoptera) from polluted areas.

The orthopteran insect Tetrix tenuicornis, collected from polluted and unpolluted areas, was used to study heavy metal accumulation and its impact on stress protein levels and on changes in the number and morphology of chromosomes in mitotic and meiotic cells. During two consecutive years, insects were collected from polluted areas of zinc-lead mine spoils near Boleslaw (Poland) and from unpolluted areas near Busko and Staszów (Poland). T. tenuicornis from the polluted area showed 1.5, 4.03, 4.32 and 41.73 times higher concentrations of copper (Cu), zinc (Zn), lead (Pb) and cadmium (Cd), respectively, than insects of the same species collected from unpolluted areas. Insects exposed to heavy metals showed only small changes, and rather a decrease in the concentration of constitutive and inducible heat shock proteins Hsp70, the level of which increases under stress conditions. A cytogenetic study of T. tenuicornis revealed intra-population anomalies in chromosome number and morphology in mitotic and meiotic cells and the presence of an additional B chromosome in germinal cells. In 50% of females collected from polluted areas, mosaic oogonial mitotic chromosome sets and diploid, hypo- or hypertetraploid, tetraploid, and octoploid chromosome numbers were detected. In turn, 14.6% of males showed a heterozygous deficiency of chromatin in L2 and M3 bivalents in addition to the presence of B chromosomes.

Multiregional origin of B chromosomes in the grasshopper Eyprepocnemis plorans.

Analysis of chromosome localization of three molecular markers, 18S-5.8S-28S rDNA, 5S rDNA and a 180 bp satDNA, showed that B chromosomes in the grasshopper Eyprepocnemis plorans originated independently in Eastern (Caucasus) and Western (Spain and Morocco) populations. Eastern B chromosomes are most likely derived from the smallest autosome, which is the only A chromosome carrying the three markers, in coincidence with Caucasian B chromosomes. Western B chromosomes, however, lack 5S rDNA and are most likely derived from the X chromosome, which is the only A chromosome carrying the two remaining markers, always in the same order with respect to the centromere, as the B chromosome.

Comparative FISH analysis in five species of Eyprepocnemidine grasshoppers.

The chromosomal localization of ribosomal DNA, and a 180 bp satellite DNA isolated from Spanish Eyprepocnemis plorans specimens, has been analysed in five Eyprepocnemidinae species collected in Russia and Central Asia. Caucasian E. plorans individuals carried each of the two DNAs, but the rDNA was limited to only two chromosomes (S(9) and S(11)) in sharp contrast to Spanish specimens that show 4-8 rDNA clusters and to Moroccan specimens which carry rDNA in almost all chromosomes. The four remaining species, however, lacked the 180 bp tandem repeat, and showed rDNA clusters in one (S(9) in Thisoicetrinus pterostichus), two (S(9) and S(10) in Eyprepocnemis unicolor; M(8) and S(11) in Heteracris adspersa), or three (S(9), S(10), and S(11) in Shirakiacris shirakii) chromosome pairs. The implications of these findings for the evolution of these two chromosome markers in this group of species are discussed.

B chromosomes, translocation between B and autosomes, and C-heterochromatin polymorphism of the grasshopper Podisma sapporensis Shir. (Orthoptera, Acrididae) in Hokkaido, northern Japan.

Seven categories of B chromosomes found in the brachypterus grasshopper Podisma sapporensis from Hokkaido populations differ in structure, size, and C-band content. The interchange between B and one autosome from M3 and sporadically M7 was observed in most of the populations examined. Such an interaction between standard and non-standard chromosomal set provides an insight into the integration of supernumerary chromosome. In addition, C-heterochromatin polymorphism was also identified in male karyotypes in some populations. These facts indicate P. sapporensis is a highly polymorphic species from the cytogenetic point of view.

Chromosome polymorphism and C-banding variation of the brachypterous grasshopper Podisma sapporensis Shir. (Orthoptera, Acrididae) in Hokkaido, northern Japan.

The grasshopper Podisma sapporensis consists of two main chromosome races in Hokkaido. The western group of populations of P. sapporensis, belonging to the XO race, has a diploid number of chromosomes 2n = 23 in the male and 2n = 24 in the female (sex determination XO male/XX female). The eastern group of populations of this species, belonging to the XY race, differs from the western one as a result of Robertsonian translocation between the originally acrocentric X chromosome and M5 autosome in homozygous state, having resulted in the forming of chromosome sex determination neo-XY male/neo-XX female (2n = 22). These races are geographically isolated by the mountainous system consisting of the Mts Daisetsu and Hidaka range, occupying the central part of the island. The hybrid zones between the races have not so far been discovered. Various levels of polymorphism for the pericentric inversions and C-banding variation exist in different chromosomes throughout populations in both chromosome races. In some solitary populations (the population at the summit of Mt Yotei, populations in the vicinity of Naganuma, Oketo, and Tanno) pericentric inversions are fixed in some pairs of chromosomes, which enables marking of the discrete karyomorphes. In the Mt Daisengen population all chromosomes are two-armed as a result of fixing the pericentric inversions. These facts contradict karyotypical conservatism of the tribe Podismini. The level of diversity of P. sapporensis karyotypes could provide a new perspective on the evolutionary process of different karyotype in Orthoptera. The considerable occurrence of polymorphism in chromosomes suggests that karyotypic diversification is undergoing in P. sapporensis. The authors also proposed that P. sapporensis would be divided into four chromosome subraces in the XO chromosome race and two chromosome subraces in the XY race, on the basis of karyotypic features. These races may have been established by fundamental climatic changes during the glacial epoch.

Cytotaxonomic characteristics of the genus Glyptotendipes Kieffer (Chironomidae, Diptera) from fish and retention ponds (Silesia, southern Poland).

The cytotaxonomic characteristics of species of the genus Glyptotendipes (Chironomidae): G. glaucus Meigen, G. paripes Edwards, and G. barbipes (Staeger) are described. The studied material was collected from a fish pond at Golysz and a sewage retention pond at Chybie in Silesia. All the studied species have the chromosome set 2n = 8, but with many specific structural and functional changes. The cytogenetic data of G. glaucus showed that the studied specimens of this species have been produced by the introgressive hybridization of two sibling species: G. glaucus x G. pallens, and subsequent crossing-over in the hybrid chromosome CD. Owing to this process, the band pattern of chromosome arm D coincided with those of G. pallens. New aberrations (homo- and heterozygous deletions in arm G as well as heterozygous inversions in chromosome arm B) were detected in G. glaucus. Specific band sequences were discovered in chromosomes of G. paripes. The band patterns established in chromosomes AB and G of this species were identical with those of Siberian populations. The banding patterns of the polytene chromosomes of G. barbipes do not differ from the standard. However, high frequency of pericentric inversion of chromosome AB was established. Many new puffs were found in the polytene chromosomes of all the studied species. Their nucleolar organizer was very sensitive to environmental changes. In G. glaucus it appeared in three different states: very active, slightly active, and heterozygous state. The organic pollution existing in the sewage retention pond may contribute to possible mutations and chromosomal damage in Glypotendipes species. Structural and functional rearrangements of the polytene chromosomes of these species mobilized their genomes and provide for survival under polluted conditions.

C-banded karyotypes of some dragonfly species from Russia. II. The families Cordulegasteridae, Corduliidae and Gomphidae.

The C-stained karyotypes of five species of three dragonfly families from Western Siberia and Kunashir Island have been analysed. Gomphus epophtalmus Sel., G. vulgatissimus (L.), Nihonogomphus ruptus (Sel. et Hag.) (Gomphidae), and Anotogaster sieboldii (Sel.) (Cordulegasteridae) showed usual character of C-heterochromatin distribution, all chromosomes have terminal C-bands. Somatochlora graeseri Sel. (Corduliidae) has unique for dragonflies type of terminal C-blocks on autosomes. Three pairs of autosomes have the very large heterochromatic blocks, other chromosomes, including the X, have no C-band.

Some aspects of karyotype of Liarina (Orthoptera: Tettigoniidae, Agraeciini) from Vietnam.

Karyotypes (chromosome number and shape) of four species of the subtribe Liarina were studied. The chromosome numbers and NF (Fundamental Number) in this group of species range from 2n Male = 33 (34) to 27(30): Liaromorpha buonluoiensis 2n Male = 33 (34), Sialaiana transiens 2n Male = 29 (34), Liara tramlapensis 2n Male = 29 (32), and Anelytra (Perianelytra) propria 2n Male = 27 (30). Cyto-taxonomy analysis indicates an intensive karyotype evolution among species belonging to three different groups of the genera. Differences of karyotypes are connected with Robertsonian fusion and tandem fusion in autosomes. Additionally, C-banding distribution and location of the NORs were studied.

Chromosome evolution in the genus Poecilimon (Orthoptera, Tettigonioidea, Phaneropteridae).

Cytotaxonomic analysis of 20 species and subspecies of the genus Poecilimon using C-banding pattern, chiasma frequency, and morphometric characteristics of the chromosomes were described. Using a cladistic analysis the chromosome data provided a basis to produce a phylogenetic tree which was compared with a tree based on morphological characters and DNA sequence data. There are important differences in the grouping of data sets to species obtained on the basis of morphology/DNA analyses and that based on chromosomes. The explanation of the differences between C-banding patterns and taxonomic proximity is probably that the C-banding pattern changes quickly as the result of the high degree of variation of constitutive heterochromatin.

Chromosome aberrations in a natural population of Tetrix tenuicornis (Sahlb.) (Tetrigidae: Orthoptera).

In the natural population of Tetrix tenuicornis from the Kraków-Czestochowa Upland 26.7% of males showed various aberrations of the morphology and behaviour of chromosomes. The high frequency of chromosome aberrations found in this population indicates their genetic instability.